The emergence of deep learning, particularly AlphaFold, has revolutionized static protein structure prediction, marking a transformative milestone in structural biology. However, protein function is not solely determined by static three-dimensional structures but is fundamentally governed by dynamic transitions between multiple conformational states. This shift from static to multi-state representations is crucial for understanding the mechanistic basis of protein function and regulation. This review outlines the fundamental concepts of protein dynamic conformations, surveys recent computational advances in modeling these dynamics in the post-AlphaFold era, and highlights key challenges, including data limitations, methodological constraints, and evaluation metrics. We also discuss potential strategies to address these challenges and explore future research directions to deepen our understanding of protein dynamics and their functional implications. This work aims to provide insights and perspectives to facilitate the ongoing development of protein conformation studies in the era of artificial intelligence-driven structural biology.

BACKGROUND: Association and cooperation among structural domains play an important role in protein function and drug design. Despite remarkable advancements in highly accurate single-domain protein structure prediction through the collaborative efforts of the community using deep learning, challenges still exist in predicting multi-domain protein structures when the evolutionary signal for a given domain pair is weak or the protein structure is large. RESULTS: To alleviate the above challenges, we proposed M-DeepAssembly, a protocol based on multi-objective protein conformation sampling algorithm for multi-domain protein structure prediction. Firstly, the inter-domain interactions and full-length sequence distance features are extracted through DeepAssembly and AlphaFold2, respectively. Secondly, subject to these features, we constructed a multi-objective energy model and designed a sampling algorithm for exploring and exploiting conformational space to generate ensembles. Finally, the output protein structure was selected from the ensembles using our in-house developed model quality assessment algorithm. On the test set of 164 multi-domain proteins, the results show that the average TM-score of M-DeepAssembly is 15.4% and 2.0% higher than AlphaFold2 and DeepAssembly, respectively. It is worth noting that there are models with higher accuracy in ensembles, achieving an improvement of 20.3% and 6.4% relative to the two baseline methods, although these models were not selected. Furthermore, when compared to the prediction results of AlphaFold2 for CASP15 multi-domain targets, M-DeepAssembly demonstrates certain performance advantages. CONCLUSIONS: M-DeepAssembly provides a distinctive multi-domain protein assembly algorithm, which can alleviate the current challenges of weak evolutionary signals and large structures to some extent by forming diverse ensembles using multi-objective protein conformation sampling algorithm. The proposed method contributes to exploring the functions of multi-domain proteins, especially providing new insights into targets with multiple conformational states.

Accurate de novo protein structure prediction remains a fundamental challenge, particularly in cases where homologous templates are unavailable or evolutionary information is weak. While end-to-end methods such as AlphaFold2 have achieved unprecedented accuracy, their closed box nature provides limited insight into the folding process and offers little flexibility for incorporating external evaluation. Here, we investigate whether model quality assessment (MQA) can be integrated into the structure prediction pipeline as a closed-loop feedback mechanism to iteratively improve prediction accuracy. In this study, we propose DGMFold, a de novo protein structure prediction method that establishes a feedback loop among three components: the geometric constraint prediction network (GeomNet), the structural simulation module, and the model quality evaluation network (EmaNet). In GeomNet, co-evolutionary features extracted from multiple sequence alignments (MSAs) are fed into an improved residual neural network to predict inter-residue geometric constraints, which are then used to guide structure folding. EmaNet then extracts 1D and 2D features from the folded structure model and employs a deep residual neural network to estimate the inter-residue distance deviation and per-residue lDDT. These evaluations are subsequently fed back into GeomNet as dynamic features, enabling iterative refinement of the predicted geometries and overall model accuracy. DGMFold was tested on 437 benchmark proteins and 20 FM targets of CASP14. Experimental results demonstrate that the closed-loop feedback mechanism significantly contributes to the performance of DGMFold, and the prediction accuracy of DGMFold outperforms that of the state-of-the-art de novo methods trRosetta and RaptorX at the time. When evaluated on the 124 human proteins for which AlphaFold2 yields TM-scores below 0.9, DGMFold achieves higher prediction accuracy than AlphaFold2 and RoseTTAFold on 71 and 72 targets, respectively, and outperforms both on 58 proteins.

Model quality evaluation is a crucial part of protein structural biology. How to distinguish high-quality models from low-quality models, and to assess which high-quality models have relatively incorrect regions for improvement, are remain a challenge. More importantly, the quality assessment of multimer models is a hot topic for structure prediction. In this study, we propose GraphCPLMQA, a novel approach for evaluating residue-level model quality that combines graph coupled networks and embeddings from protein language models. The GraphCPLMQA consists of a graph encoding module and a transform-based convolutional decoding module. In encoding module, the underlying relational representations of sequence and high-dimensional geometry structure are extracted by protein language models with Evolutionary Scale Modeling. In decoding module, the mapping connection between structure and quality is inferred by the representations and low-dimensional features. Specifically, the triangular location and residue level contact order features are designed to enhance the association between the local structure and the overall topology. Experimental results demonstrate that GraphCPLMQA using single-sequence embedding achieves the best performance compared with the CASP15 residue-level interface evaluation methods among 9108 models in the local residue interface test set of CASP15 multimers. In CAMEO blind test (20 May 2022 to 13 August 2022), GraphCPLMQA ranked first compared with other servers (https://www.cameo3d.org/quality-estimation). GraphCPLMQA also outperforms state-of-the-art methods on 19, 035 models in CASP13 and CASP14 monomer test set.

Protein sequence design can provide valuable insights into biopharmaceuticals and disease treatments. Currently, most protein sequence design methods based on deep learning focus on network architecture optimization, while ignoring protein-specific physicochemical features. Inspired by the successful application of structure templates and pre-trained models in the protein structure prediction, we explored whether the representation of structural sequence profile can be used for protein sequence design. In this work, we propose SPDesign, a method for protein sequence design based on structural sequence profile using ultrafast shape recognition. Given an input backbone structure, SPDesign utilizes ultrafast shape recognition vectors to accelerate the search for similar protein structures in our in-house PAcluster80 structure database and then extracts the sequence profile through structure alignment. Combined with structural pre-trained knowledge and geometric features, they are further fed into an enhanced graph neural network for sequence prediction. The results show that SPDesign significantly outperforms the state-of-the-art methods, such as ProteinMPNN, Pifold and LM-Design, leading to 21.89%, 15.54% and 11.4% accuracy gains in sequence recovery rate on CATH 4.2 benchmark, respectively. Encouraging results also have been achieved on orphan and de novo (designed) benchmarks with few homologous sequences. Furthermore, analysis conducted by the PDBench tool suggests that SPDesign performs well in subdivided structures. More interestingly, we found that SPDesign can well reconstruct the sequences of some proteins that have similar structures but different sequences. Finally, the structural modeling verification experiment indicates that the sequences designed by SPDesign can fold into the native structures more accurately.

The breakthrough in cryo-electron microscopy (cryo-EM) technology has led to an increasing number of density maps of biological macromolecules. However, constructing accurate protein complex atomic structures from cryo-EM maps remains a challenge. In this study, we extend our previously developed DEMO-EM to present DEMO-EM2, an automated method for constructing protein complex models from cryo-EM maps through an iterative assembly procedure intertwining chain- and domain-level matching and fitting for predicted chain models. The method was carefully evaluated on 27 cryo-electron tomography (cryo-ET) maps and 16 single-particle EM maps, where DEMO-EM2 models achieved an average TM-score of 0.92, outperforming those of state-of-the-art methods. The results demonstrate an efficient method that enables the rapid and reliable solution of challenging cryo-EM structure modeling problems.

AlphaFold2 has achieved a major breakthrough in end-to-end prediction for static protein structures. However, protein conformational change is considered to be a key factor in protein biological function. Inter-residue multiple distances prediction is of great significance for research on protein multiple conformations exploration. In this study, we proposed an inter-residue multiple distances prediction method, DeepMDisPre, based on an improved network which integrates triangle update, axial attention and ResNet to predict multiple distances of residue pairs. We built a dataset which contains proteins with a single structure and proteins with multiple conformations to train the network. We tested DeepMDisPre on 114 proteins with multiple conformations. The results show that the inter-residue distance distribution predicted by DeepMDisPre tends to have multiple peaks for flexible residue pairs than for rigid residue pairs. On two cases of proteins with multiple conformations, we modeled the multiple conformations relatively accurately by using the predicted inter-residue multiple distances. In addition, we also tested the performance of DeepMDisPre on 279 proteins with a single structure. Experimental results demonstrate that the average contact accuracy of DeepMDisPre is higher than that of the comparative method. In terms of static protein modeling, the average TM-score of the 3D models built by DeepMDisPre is also improved compared with the comparative method.

AlphaFold2 achieved a breakthrough in protein structure prediction through the end-to-end deep learning method, which can predict nearly all single-domain proteins at experimental resolution. However, the prediction accuracy of full-chain proteins is generally lower than that of single-domain proteins because of the incorrect interactions between domains. In this work, we develop an inter-domain distance prediction method, named DeepIDDP. In DeepIDDP, we design a neural network with attention mechanisms, where two new inter-domain features are used to enhance the ability to capture the interactions between domains. Furthermore, we propose a data enhancement strategy termed DPMSA, which is employed to deal with the absence of co-evolutionary information on targets. We integrate DeepIDDP into our previously developed domain assembly method SADA, termed SADA-DeepIDDP. Tested on a given multi-domain benchmark dataset, the accuracy of SADA-DeepIDDP inter-domain distance prediction is 11.3% and 21.6% higher than trRosettaX and trRosetta, respectively. The accuracy of the domain assembly model is 2.5% higher than that of SADA. Meanwhile, we reassemble 68 human multi-domain protein models with TM-score ≤ 0.80 from the AlphaFold protein structure database, where the average TM-score is improved by 11.8% after the reassembly by our method. The online server is at http://zhanglab-bioinf.com/DeepIDDP/.

The scoring models used for protein structure modeling and ranking are mainly divided into unified field and protein-specific scoring functions. Although protein structure prediction has made tremendous progress since CASP14, the modeling accuracy still cannot meet the requirements to a certain extent. Especially, accurate modeling of multi-domain and orphan proteins remains a challenge. Therefore, an accurate and efficient protein scoring model should be developed urgently to guide the protein structure folding or ranking through deep learning. In this work, we propose a protein structure global scoring model based on equivariant graph neural network (EGNN), named GraphGPSM, to guide protein structure modeling and ranking. We construct an EGNN architecture, and a message passing mechanism is designed to update and transmit information between nodes and edges of the graph. Finally, the global score of the protein model is output through a multilayer perceptron. Residue-level ultrafast shape recognition is used to describe the relationship between residues and the overall structure topology, and distance and direction encoded by Gaussian radial basis functions are designed to represent the overall topology of the protein backbone. These two features are combined with Rosetta energy terms, backbone dihedral angles and inter-residue distance and orientations to represent the protein model and embedded into the nodes and edges of the graph neural network. The experimental results on the CASP13, CASP14 and CAMEO test sets show that the scores of our developed GraphGPSM have a strong correlation with the TM-score of the models, which are significantly better than those of the unified field score function REF2015 and the state-of-the-art local lDDT-based scoring models ModFOLD8, ProQ3D and DeepAccNet, etc. The modeling experimental results on 484 test proteins demonstrate that GraphGPSM can greatly improve the modeling accuracy. GraphGPSM is further used to model 35 orphan proteins and 57 multi-domain proteins. The results show that the average TM-score of the models predicted by GraphGPSM is 13.2 and 7.1% higher than that of the models predicted by AlphaFold2. GraphGPSM also participates in CASP15 and achieves competitive performance in global accuracy estimation.

Protein model quality assessment plays an important role in protein structure prediction, protein design and drug discovery. In this work, DeepUMQA2, a substantially improved version of DeepUMQA for protein model quality assessment, is proposed. First, sequence features containing protein co-evolution information and structural features reflecting family information are extracted to complement model-dependent features. Second, a novel backbone network based on triangular multiplication update and axial attention mechanism is designed to enhance information exchange between inter-residue pairs. On CASP13 and CASP14 datasets, the performance of DeepUMQA2 increases by 20.5 and 20.4% compared with DeepUMQA, respectively (measured by top 1 loss). Moreover, on the three-month CAMEO dataset (11 March to 04 June 2022), DeepUMQA2 outperforms DeepUMQA by 15.5% (measured by local AUC0,0.2) and ranks first among all competing server methods in CAMEO blind test. Experimental results show that DeepUMQA2 outperforms state-of-the-art model quality assessment methods, such as ProQ3D-LDDT, ModFOLD8, and DeepAccNet and DeepUMQA2 can select more suitable best models than state-of-the-art protein structure methods, such as AlphaFold2, RoseTTAFold and I-TASSER, provided themselves.

MOTIVATION: Model quality assessment is a crucial part of protein structure prediction and a gateway to proper usage of models in biomedical applications. Many methods have been proposed for assessing the quality of structural models of protein monomers, but few methods for evaluating protein complex models. As protein complex structure prediction becomes a new challenge, there is an urgent need for model quality assessment methods that can accurately assess the accuracy of interface residues of complex structures. RESULTS: Here, we present DeepUMQA3, a web server for evaluating the accuracy of interface residues of protein complex structures using deep neural networks. For an input complex structure, features are extracted from three levels of overall complex, intra-monomer, and inter-monomer, and an improved deep residual neural network is used to predict per-residue lDDT and interface residue accuracy. DeepUMQA3 ranks first in the blind test of interface residue accuracy estimation in CASP15, with Pearson, Spearman, and AUC of 0.564, 0.535, and 0.755 under the lDDT measurement, which are 17.6%, 23.6%, and 10.9% higher than the second best method, respectively. DeepUMQA3 can also assess the accuracy of all residues in the entire complex and distinguish high- and low-precision residues. AVAILABILITY AND IMPLEMENTATION: The web sever of DeepUMQA3 are freely available at http://zhanglab-bioinf.com/DeepUMQA_server/.

The study of protein folding mechanism is a challenge in molecular biology, which is of great significance for revealing the movement rules of biological macromolecules, understanding the pathogenic mechanism of folding diseases, and designing protein engineering materials. Based on the hypothesis that the conformational sampling trajectory contain the information of folding pathway, we propose a protein folding pathway prediction algorithm named Pathfinder. Firstly, Pathfinder performs large-scale sampling of the conformational space and clusters the decoys obtained in the sampling. The heterogeneous conformations obtained by clustering are named seed states. Then, a resampling algorithm that is not constrained by the local energy basin is designed to obtain the transition probabilities of seed states. Finally, protein folding pathways are inferred from the maximum transition probabilities of seed states. The proposed Pathfinder is tested on our developed test set (34 proteins). For 11 widely studied proteins, we correctly predicted their folding pathways and specifically analyzed 5 of them. For 13 proteins, we predicted their folding pathways to be further verified by biological experiments. For 6 proteins, we analyzed the reasons for the low prediction accuracy. For the other 4 proteins without biological experiment results, potential folding pathways were predicted to provide new insights into protein folding mechanism. The results reveal that structural analogs may have different folding pathways to express different biological functions, homologous proteins may contain common folding pathways, and α-helices may be more prone to early protein folding than β-strands.

Domain boundary prediction is one of the most important problems in the study of protein structure and function, especially for large proteins. At present, most domain boundary prediction methods have low accuracy and limitations in dealing with multi-domain proteins. In this study, we develop a sequence-based protein domain boundary prediction, named DomBpred. In DomBpred, the input sequence is first classified as either a single-domain protein or a multi-domain protein through a designed effective sequence metric based on a constructed single-domain sequence library. For the multi-domain protein, a domain-residue clustering algorithm inspired by Ising model is proposed to cluster the spatially close residues according inter-residue distance. The unclassified residues and the residues at the edge of the cluster are then tuned by the secondary structure to form potential cut points. Finally, a domain boundary scoring function is proposed to recursively evaluate the potential cut points to generate the domain boundary. DomBpred is tested on a large-scale test set of FUpred comprising 2549 proteins. Experimental results show that DomBpred better performs than the state-of-the-art methods in classifying whether protein sequences are composed by single or multiple domains, and the Matthew's correlation coefficient is 0.882. Moreover, on 849 multi-domain proteins, the domain boundary distance and normalised domain overlap scores of DomBpred are 0.523 and 0.824, respectively, which are 5.0% and 4.2% higher than those of the best comparison method, respectively. Comparison with other methods on the given test set shows that DomBpred outperforms most state-of-the-art sequence-based methods and even achieves better results than the top-level template-based method. The executable program is freely available at https://github.com/iobio-zjut/DomBpred and the online server at http://zhanglab-bioinf.com/DomBpred/.

Although remarkable achievements, such as AlphaFold2, have been made in end-to-end structure prediction, fragment libraries remain essential for de novo protein structure prediction, which can help explore and understand the protein-folding mechanism. In this work, we developed a variable-length fragment library (VFlib). In VFlib, a master structure database was first constructed from the Protein Data Bank through sequence clustering. The hidden Markov model (HMM) profile of each protein in the master structure database was generated by HHsuite, and the secondary structure of each protein was calculated by DSSP. For the query sequence, the HMM-profile was first constructed. Then, variable-length fragments were retrieved from the master structure database through dynamically variable-length profile-profile comparison. A complete method for chopping the query HMM-profile during this process was proposed to obtain fragments with increased diversity. Finally, secondary structure information was used to further screen the retrieved fragments to generate the final fragment library of specific query sequence. The experimental results obtained with a set of 120 nonredundant proteins show that the global precision and coverage of the fragment library generated by VFlib were 55.04% and 94.95% at the RMSD cutoff of 1.5 Å, respectively. Compared with the benchmark method of NNMake, the global precision of our fragment library had increased by 62.89% with equivalent coverage. Furthermore, the fragments generated by VFlib and NNMake were used to predict structure models through fragment assembly. Controlled experimental results demonstrate that the average TM-score of VFlib was 16.00% higher than that of NNMake.

Meta contact, which combines different contact maps into one to improve contact prediction accuracy and effectively reduce the noise from a single contact map, is a widely used method. However, protein structure prediction using meta contact cannot fully exploit the information carried by original contact maps. In this work, a multi contact-based folding method under the evolutionary algorithm framework, MultiCFold, is proposed. In MultiCFold, the thorough information of different contact maps is directly used by populations to guide protein structure folding. In addition, noncontact is considered as an effective supplement to contact information and can further assist protein folding. MultiCFold is tested on a set of 120 nonredundant proteins, and the average TM-score and average RMSD reach 0.617 and 5.815 Å, respectively. Compared with the meta contact-based method, MetaCFold, average TM-score and average RMSD have a 6.62 and 8.82% improvement. In particular, the import of noncontact information increases the average TM-score by 6.30%. Furthermore, MultiCFold is compared with four state-of-the-art methods of CASP13 on the 24 FM targets, and results show that MultiCFold is significantly better than other methods after the full-atom relax procedure.

With the rapid progress of deep learning in cryo-electron microscopy and protein structure prediction, improving the accuracy of the protein structure model by using a density map and predicted contact/distance map through deep learning has become an urgent need for robust methods. Thus, designing an effective protein structure optimization strategy based on the density map and predicted contact/distance map is critical to improving the accuracy of structure refinement. In this article, a protein structure optimization method based on the density map and predicted contact/distance map by deep-learning technology was proposed in accordance with the result of matching between the density map and the initial model. Physics- and knowledge-based energy functions, integrated with Cryo-EM density map data and deep-learning data, were used to optimize the protein structure in the simulation. The dynamic confidence score was introduced to the iterative process for choosing whether it is a density map or a contact/distance map to dominate the movement in the simulation to improve the accuracy of refinement. The protocol was tested on a large set of 224 non-homologous membrane proteins and generated 214 structural models with correct folds, where 4.5% of structural models were generated from structural models with incorrect folds. Compared with other state-of-the-art methods, the major advantage of the proposed methods lies in the skills for using density map and contact/distance map in the simulation, as well as the new energy function in the re-assembly simulations. Overall, the results demonstrated that this strategy is a valuable approach and ready to use for atomic-level structure refinement using cryo-EM density map and predicted contact/distance map.

MOTIVATION: Protein model quality assessment is a key component of protein structure prediction. In recent research, the voxelization feature was used to characterize the local structural information of residues, but it may be insufficient for describing residue-level topological information. Design features that can further reflect residue-level topology when combined with deep learning methods are therefore crucial to improve the performance of model quality assessment. RESULTS: We developed a deep-learning method, DeepUMQA, based on Ultrafast Shape Recognition (USR) for the residue-level single-model quality assessment. In the framework of the deep residual neural network, the residue-level USR feature was introduced to describe the topological relationship between the residue and overall structure by calculating the first moment of a set of residue distance sets and then combined with 1D, 2D and voxelization features to assess the quality of the model. Experimental results on the CASP13, CASP14 test datasets and CAMEO blind test show that USR could supplement the voxelization features to comprehensively characterize residue structure information and significantly improve model assessment accuracy. The performance of DeepUMQA ranks among the top during the state-of-the-art single-model quality assessment methods, including ProQ2, ProQ3, ProQ3D, Ornate, VoroMQA, ProteinGCN, ResNetQA, QDeep, GraphQA, ModFOLD6, ModFOLD7, ModFOLD8, QMEAN3, QMEANDisCo3 and DeepAccNet. AVAILABILITY AND IMPLEMENTATION: The DeepUMQA server is freely available at http://zhanglab-bioinf.com/DeepUMQA/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MOTIVATION: With the breakthrough of AlphaFold2, the protein structure prediction problem has made remarkable progress through deep learning end-to-end techniques, in which correct folds could be built for nearly all single-domain proteins. However, the full-chain modelling appears to be lower on average accuracy than that for the constituent domains and requires higher demand on computing hardware, indicating the performance of full-chain modelling still needs to be improved. In this study, we investigate whether the predicted accuracy of the full-chain model can be further improved by domain assembly assisted by deep learning. RESULTS: In this article, we developed a structural analogue-based protein structure domain assembly method assisted by deep learning, named SADA. In SADA, a multi-domain protein structure database was constructed for the full-chain analogue detection using individual domain models. Starting from the initial model constructed from the analogue, the domain assembly simulation was performed to generate the full-chain model through a two-stage differential evolution algorithm guided by the energy function with an inter-residue distance potential predicted by deep learning. SADA was compared with the state-of-the-art domain assembly methods on 356 benchmark proteins, and the average TM-score of SADA models is 8.1% and 27.0% higher than that of DEMO and AIDA, respectively. We also assembled 293 human multi-domain proteins, where the average TM-score of the full-chain model after the assembly by SADA is 1.1% higher than that of the model by AlphaFold2. To conclude, we find that the domains often interact in the similar way in the quaternary orientations if the domains have similar tertiary structures. Furthermore, homologous templates and structural analogues are complementary for multi-domain protein full-chain modelling. AVAILABILITY AND IMPLEMENTATION: http://zhanglab-bioinf.com/SADA. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MOTIVATION: Accurately identifying protein-ATP binding poses is significantly valuable for both basic structure biology and drug discovery. Although many docking methods have been designed, most of them require a user-defined binding site and are difficult to achieve a high-quality protein-ATP docking result. It is critical to develop a protein-ATP-specific blind docking method without user-defined binding sites. RESULTS: Here, we present ATPdock, a template-based method for docking ATP into protein. For each query protein, if no pocket site is given, ATPdock first identifies its most potential pocket using ATPbind, an ATP-binding site predictor; then, the template pocket, which is most similar to the given or identified pocket, is searched from the database of pocket-ligand structures using APoc, a pocket structural alignment tool; thirdly, the rough docking pose of ATP (rdATP) is generated using LS-align, a ligand structural alignment tool, to align the initial ATP pose to the template ligand corresponding to template pocket; finally, the Metropolis Monte Carlo simulation is used to fine-tune the rdATP under the guidance of AutoDock Vina energy function. Benchmark tests show that ATPdock significantly outperforms other state-of-the-art methods in docking accuracy. AVAILABILITY AND IMPLEMENTATION: https://jun-csbio.github.io/atpdock/. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Protein-DNA interactions play an important role in diverse biological processes. Accurately identifying protein-DNA binding residues is a critical but challenging task for protein function annotations and drug design. Although wet-lab experimental methods are the most accurate way to identify protein-DNA binding residues, they are time consuming and labor intensive. There is an urgent need to develop computational methods to rapidly and accurately predict protein-DNA binding residues. In this study, we propose a novel sequence-based method, named PredDBR, for predicting DNA-binding residues. In PredDBR, for each query protein, its position-specific frequency matrix (PSFM), predicted secondary structure (PSS), and predicted probabilities of ligand-binding residues (PPLBR) are first generated as three feature sources. Secondly, for each feature source, the sliding window technique is employed to extract the matrix-format feature of each residue. Then, we design two strategies, i.e., square root (SR) and average (AVE), to separately transform PSFM-based and two predicted feature source-based, i.e., PSS-based and PPLBR-based, matrix-format features of each residue into three corresponding cube-format features. Finally, after serially combining the three cube-format features, the ensemble classifier is generated via applying bagging strategy to multiple base classifiers built by the framework of 2D convolutional neural network. The computational experimental results demonstrate that the proposed PredDBR achieves an average overall accuracy of 93.7% and a Mathew's correlation coefficient of 0.405 on two independent validation datasets and outperforms several state-of-the-art sequenced-based protein-DNA binding residue predictors. The PredDBR web-server is available at https://jun-csbio.github.io/PredDBR/.

De novo protein structure prediction is a challenging problem that requires both an accurate energy function and an efficient conformation sampling method. In this study, a de novo structure prediction method, named CoDiFold, is proposed. In CoDiFold, contacts and distance profiles are organically combined into the Rosetta low-resolution energy function to improve the accuracy of energy function. As a result, the correlation between energy and root mean square deviation (RMSD) is improved. In addition, a population-based multi-mutation strategy is designed to balance the exploration and exploitation of conformation space sampling. The average RMSD of the models generated by the proposed protocol is decreased by 49.24 and 45.21 percent in the test set with 43 proteins compared with those of Rosetta and QUARK de novo protocols, respectively. The results also demonstrate that the structures predicted by proposed CoDiFold are comparable to the state-of-the-art methods for the 10 FM targets of CASP13. The source code and executable versions are freely available at http://github.com/iobio-zjut/CoDiFold.

Advances in the prediction of the inter-residue distance for a protein sequence have increased the accuracy to predict the correct folds of proteins with distance information. Here, we propose a distance-guided protein folding algorithm based on generalized descent direction, named GDDfold, which achieves effective structural perturbation and potential minimization in two stages. In the global stage, random-based direction is designed using evolutionary knowledge, which guides conformation population to cross potential barriers and explore conformational space rapidly in a large range. In the local stage, locally rugged potential landscape can be explored with the aid of conjugate-based direction integrated into a specific search strategy, which can improve the exploitation ability. GDDfold is tested on 347 proteins of a benchmark set, 24 template-free modeling (FM) approaches targets of CASP13 and 20 FM targets of CASP14. Results show that GDDfold correctly folds [template modeling (TM) score ≥ = 0.5] 316 out of 347 proteins, where 65 proteins have TM scores that are greater than 0.8, and significantly outperforms Rosetta-dist (distance-assisted fragment assembly method) and L-BFGSfold (distance geometry optimization method). On CASP FM targets, GDDfold is comparable with five state-of-the-art full-version methods, namely, Quark, RaptorX, Rosetta, MULTICOM and trRosetta in the CASP 13 and 14 server groups.

MOTIVATION: With the great progress of deep learning-based inter-residue contact/distance prediction, the discrete space formed by fragment assembly cannot satisfy the distance constraint well. Thus, the optimal solution of the continuous space may not be achieved. Designing an effective closed-loop continuous dihedral angle optimization strategy that complements the discrete fragment assembly is crucial to improve the performance of the distance-assisted fragment assembly method. RESULTS: In this article, we proposed a de novo protein structure prediction method called IPTDFold based on closed-loop iterative partition sampling, topology adjustment and residue-level distance deviation optimization. First, local dihedral angle crossover and mutation operators are designed to explore the conformational space extensively and achieve information exchange between the conformations in the population. Then, the dihedral angle rotation model of loop region with partial inter-residue distance constraints is constructed, and the rotation angle satisfying the constraints is obtained by differential evolution algorithm, so as to adjust the spatial position relationship between the secondary structures. Finally, the residue distance deviation is evaluated according to the difference between the conformation and the predicted distance, and the dihedral angle of the residue is optimized with biased probability. The final model is generated by iterating the above three steps. IPTDFold is tested on 462 benchmark proteins, 24 FM targets of CASP13 and 20 FM targets of CASP14. Results show that IPTDFold is significantly superior to the distance-assisted fragment assembly method Rosetta_D (Rosetta with distance). In particular, the prediction accuracy of IPTDFold does not decrease as the length of the protein increases. When using the same FastRelax protocol, the prediction accuracy of IPTDFold is significantly superior to that of trRosetta without orientation constraints, and is equivalent to that of the full version of trRosetta. AVAILABILITYAND IMPLEMENTATION: The source code and executable are freely available at https://github.com/iobio-zjut/IPTDFold. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MOTIVATION: Massive local minima on the protein energy landscape often cause traditional conformational sampling algorithms to be easily trapped in local basin regions, because they find it difficult to overcome high-energy barriers. Also, the lowest energy conformation may not correspond to the native structure due to the inaccuracy of energy models. This study investigates whether these two problems can be alleviated by a sequential niche technique without loss of accuracy. RESULTS: A sequential niche multimodal conformational sampling algorithm for protein structure prediction (SNfold) is proposed in this study. In SNfold, a derating function is designed based on the knowledge learned from the previous sampling and used to construct a series of sampling-guided energy functions. These functions then help the sampling algorithm overcome high-energy barriers and avoid the re-sampling of the explored regions. In inaccurate protein energy models, the high-energy conformation that may correspond to the native structure can be sampled with successively updated sampling-guided energy functions. The proposed SNfold is tested on 300 benchmark proteins, 24 CASP13 and 19 CASP14 FM targets. Results show that SNfold correctly folds (TM-score ≥ 0.5) 231 out of 300 proteins. In particular, compared with Rosetta restrained by distance (Rosetta-dist), SNfold achieves higher average TM-score and improves the sampling efficiency by more than 100 times. On several CASP FM targets, SNfold also shows good performance compared with four state-of-the-art servers in CASP. As a plug-in conformational sampling algorithm, SNfold can be extended to other protein structure prediction methods. AVAILABILITY AND IMPLEMENTATION: The source code and executable versions are freely available at https://github.com/iobio-zjut/SNfold. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MOTIVATION: The mathematically optimal solution in computational protein folding simulations does not always correspond to the native structure, due to the imperfection of the energy force fields. There is therefore a need to search for more diverse suboptimal solutions in order to identify the states close to the native. We propose a novel multimodal optimization protocol to improve the conformation sampling efficiency and modeling accuracy of de novo protein structure folding simulations. RESULTS: A distance-assisted multimodal optimization sampling algorithm, MMpred, is proposed for de novo protein structure prediction. The protocol consists of three stages: The first is a modal exploration stage, in which a structural similarity evaluation model DMscore is designed to control the diversity of conformations, generating a population of diverse structures in different low-energy basins. The second is a modal maintaining stage, where an adaptive clustering algorithm MNDcluster is proposed to divide the populations and merge the modal by adjusting the annealing temperature to locate the promising basins. In the last stage of modal exploitation, a greedy search strategy is used to accelerate the convergence of the modal. Distance constraint information is used to construct the conformation scoring model to guide sampling. MMpred is tested on a large set of 320 non-redundant proteins, where MMpred obtains models with TM-score≥0.5 on 291 cases, which is 28% higher than that of Rosetta guided with the same set of distance constraints. In addition, on 320 benchmark proteins, the enhanced version of MMpred (E-MMpred) has 167 targets better than trRosetta when the best of five models are evaluated. The average TM-score of the best model of E-MMpred is 0.732, which is comparable to trRosetta (0.730). AVAILABILITY AND IMPLEMENTATION: The source code and executable are freely available at https://github.com/iobio-zjut/MMpred. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Ab initio protein structure prediction is one of the most challenging problems in computational biology. Multistage algorithms are widely used in ab initio protein structure prediction. The different computational costs of a multistage algorithm for different proteins are important to be considered. In this study, a population-based algorithm guided by information entropy (PAIE), which includes exploration and exploitation stages, is proposed for protein structure prediction. In PAIE, an entropy-based stage switch strategy is designed to switch from the exploration stage to the exploitation stage. Torsion angle statistical information is also deduced from the first stage and employed to enhance the exploitation in the second stage. Results indicate that an improvement in the performance of protein structure prediction in a benchmark of 30 proteins and 17 other free modeling targets in CASP.

MOTIVATION: Regions that connect secondary structure elements in a protein are known as loops, whose slight change will produce dramatic effect on the entire topology. This study investigates whether the accuracy of protein structure prediction can be improved using a loop-specific sampling strategy. RESULTS: A novel de novo protein structure prediction method that combines global exploration and loop perturbation is proposed in this study. In the global exploration phase, the fragment recombination and assembly are used to explore the massive conformational space and generate native-like topology. In the loop perturbation phase, a loop-specific local perturbation model is designed to improve the accuracy of the conformation and is solved by differential evolution algorithm. These two phases enable a cooperation between global exploration and local exploitation. The filtered contact information is used to construct the conformation selection model for guiding the sampling. The proposed CGLFold is tested on 145 benchmark proteins, 14 free modeling (FM) targets of CASP13 and 29 FM targets of CASP12. The experimental results show that the loop-specific local perturbation can increase the structure diversity and success rate of conformational update and gradually improve conformation accuracy. CGLFold obtains template modeling score ≥ 0.5 models on 95 standard test proteins, 7 FM targets of CASP13 and 9 FM targets of CASP12. AVAILABILITY AND IMPLEMENTATION: The source code and executable versions are freely available at https://github.com/iobio-zjut/CGLFold. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

MOTIVATION: At present, a fundamental challenge in single-cell RNA-sequencing data analysis is functional interpretation and annotation of cell clusters. Biological pathways in distinct cell types have different activation patterns, which facilitates the understanding of cell functions using single-cell transcriptomics. However, no effective web tool has been implemented for single-cell transcriptome data analysis based on prior biological pathway knowledge. RESULTS: Here, we present scTPA, a web-based platform for pathway-based analysis of single-cell RNA-seq data in human and mouse. scTPA incorporates four widely-used gene set enrichment methods to estimate the pathway activation scores of single cells based on a collection of available biological pathways with different functional and taxonomic classifications. The clustering analysis and cell-type-specific activation pathway identification were provided for the functional interpretation of cell types from a pathway-oriented perspective. An intuitive interface allows users to conveniently visualize and download single-cell pathway signatures. Overall, scTPA is a comprehensive tool for the identification of pathway activation signatures for the analysis of single cell heterogeneity. AVAILABILITY AND IMPLEMENTATION: http://sctpa.bio-data.cn/sctpa. CONTACT: [email protected] or [email protected] or [email protected]. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Accurately identifying DNA-binding proteins (DBPs) from protein sequence information is an important but challenging task for protein function annotations. In this paper, we establish a novel computational method, named TargetDBP, for accurately targeting DBPs from primary sequences. In TargetDBP, four single-view features, i.e., AAC (Amino Acid Composition), PsePSSM (Pseudo Position-Specific Scoring Matrix), PsePRSA (Pseudo Predicted Relative Solvent Accessibility), and PsePPDBS (Pseudo Predicted Probabilities of DNA-Binding Sites), are first extracted to represent different base features, respectively. Second, differential evolution algorithm is employed to learn the weights of four base features. Using the learned weights, we weightedly combine these base features to form the original super feature. An excellent subset of the super feature is then selected by using a suitable feature selection algorithm SVM-REF+CBR (Support Vector Machine Recursive Feature Elimination with Correlation Bias Reduction). Finally, the prediction model is learned via using support vector machine on the selected feature subset. We also construct a new gold-standard and non-redundant benchmark dataset from PDB database to evaluate and compare the proposed TargetDBP with other existing predictors. On this new dataset, TargetDBP can achieve higher performance than other state-of-the-art predictors. The TargetDBP web server and datasets are freely available at http://csbio.njust.edu.cn/bioinf/targetdbp/ for academic use.

Ab initio protein tertiary structure prediction is one of the long-standing problems in structural bioinformatics. With the help of residue-residue contact and secondary structure prediction information, the accuracy of ab initio structure prediction can be enhanced. In this study, an improved differential evolution with secondary structure and residue-residue contact information referred to as SCDE is proposed for protein structure prediction. In SCDE, two score models based on secondary structure and contact information are proposed, and two selection strategies, namely, secondary structure-based selection strategy and contact-based selection strategy, are designed to guide conformation space search. A probability distribution function is designed to balance these two selection strategies. Experimental results on a benchmark dataset with 28 proteins and four free model targets in CASP12 demonstrate that the proposed SCDE is effective and efficient.

De novo protein structure prediction can be treated as a conformational space optimization problem under the guidance of an energy function. However, it is a challenge of how to design an accurate energy function which ensures low-energy conformations close to native structures. Fortunately, recent studies have shown that the accuracy of de novo protein structure prediction can be significantly improved by integrating the residue-residue distance information. In this paper, a two-stage distance feature-based optimization algorithm (TDFO) for de novo protein structure prediction is proposed within the framework of evolutionary algorithm. In TDFO, a similarity model is first designed by using feature information which is extracted from distance profiles by bisecting K-means algorithm. The similarity model-based selection strategy is then developed to guide conformation search, and thus improve the quality of the predicted models. Moreover, global and local mutation strategies are designed, and a state estimation strategy is also proposed to strike a trade-off between the exploration and exploitation of the search space. Experimental results of 35 benchmark proteins show that the proposed TDFO can improve prediction accuracy for a large portion of test proteins.

Various mutation strategies show distinct advantages in differential evolution (DE). The cooperation of multiple strategies in the evolutionary process may be effective. This paper presents an underestimation-assisted global and local cooperative DE to simultaneously enhance the effectiveness and efficiency. In the proposed algorithm, two phases, namely, the global exploration and the local exploitation, are performed in each generation. In the global phase, a set of trial vectors is produced for each target individual by employing multiple strategies with strong exploration capability. Afterward, an adaptive underestimation model with a self-adapted slope control parameter is proposed to evaluate these trial vectors, the best of which is selected as the candidate. In the local phase, the better-based strategies guided by individuals that are better than the target individual are designed. For each individual accepted in the global phase, multiple trial vectors are generated by using these strategies and filtered by the underestimation value. The cooperation between the global and local phases includes two aspects. First, both of them concentrate on generating better individuals for the next generation. Second, the global phase aims to locate promising regions quickly while the local phase serves as a local search for enhancing convergence. Moreover, a simple mechanism is designed to determine the parameter of DE adaptively in the searching process. Finally, the proposed approach is applied to predict the protein 3D structure. Experimental studies on classical benchmark functions, CEC test sets, and protein structure prediction problem show that the proposed approach is superior to the competitors.

With the improvement of living standards, people pay more attention to health, which is significant to analyze people's hospitalizing behaviors. The wide use of mobile devices generates a great deal of data, which contains a lot of travel information about residents. Many people would like to see a doctor through calling an online car hailing for its convenience. Thus, based on big trajectory data generated by the online car hailing, the hospitalizing behaviors of residents are analyzed in this paper. The hospitalizing behaviors are analyzed from two aspects. One is performed from the temporal aspect, in which the daily numbers of trips of hospitalizing behaviors under different modes are analyzed. The other one is performed from the spatial aspect, in which the hot hospitals, popularity, and gravity distribution of hospitals are analyzed. Based on the spatial analysis, the network constructed by the hot hospitals is also analyzed. The results show that the hospitalizing behavior analysis can reflect the hospitalizing behaviors in detail, which can make contributions to the decision-making of infrastructure configuration for institutions, such as urban planning departments and hospitals.

As we know, the performance of differential evolution (DE) highly depends on the mutation strategy. However, it is difficult to choose a suitable mutation strategy for a specific problem or different running stages. This paper proposes an underestimation-based multimutation strategy (UMS) for DE. In the UMS, a set of candidate offsprings are simultaneously generated for each target individual by utilizing multiple mutation strategies. Then a cheap abstract convex underestimation model is built based on some selected individuals to obtain the underestimation value of each candidate offspring. According to the quality of each candidate offspring measured by the underestimation value, the most promising candidate solution is chosen as the offspring. Compared to the existing probability-based multimutation techniques, no mutation strategies are lost during the search process as each mutation strategy has the same probability to generate a candidate solution. Moreover, no extra function evaluations are produced because the candidate solutions are filtered by the underestimation value. The UMS is integrated into some DE variants and compared with their original algorithms and several advanced DE approaches over the CEC 2013 and 2014 benchmark sets. Additionally, a well-known real-world problem is employed to evaluate the performance of the UMS. Experimental results show that the proposed UMS can improve the performance of the advanced DE variants.

De novo protein structure prediction aims to search for low-energy conformations as it follows the thermodynamics hypothesis that places native conformations at the global minimum of the protein energy surface. However, the native conformation is not necessarily located in the lowest-energy regions owing to the inaccuracies of the energy model. This study presents a differential evolution algorithm using distance profile-based selection strategy to sample conformations with reasonable structure effectively. In the proposed algorithm, besides energy, the residue-residue distance is considered another measure of the conformation. The average distance errors of decoys between the distance of each residue pair and the corresponding distance in the distance profiles are first calculated when the trial conformation yields a larger energy value than that of the target. Then, the distance acceptance probability of the trial conformation is designed based on distance profiles if the trial conformation obtains a lower average distance error compared with that of the target conformation. The trial conformation is accepted to the next generation in accordance with its distance acceptance probability. By using the dual constraints of energy and distance in guiding sampling, the algorithm can sample conformations with lower energies and more reasonable structures. Experimental results of 28 benchmark proteins show that the proposed algorithm can effectively predict near-native protein structures.

In differential evolution (DE), different strategies applied in different evolutionary stages may be more effective than a single strategy used in the entire evolutionary process. However, it is not trivial to appropriately determine the evolutionary stage. In this paper, we present an abstract convex underestimation-assisted multistage DE. In the proposed algorithm, the underestimation is calculated through the supporting vectors of some neighboring individuals. Based on the variation of the average underestimation error (UE), the evolutionary process is divided into three stages. Each stage includes a pool of suitable candidate strategies. At the beginning of each generation, the evolutionary stage is first estimated according to the average UE of the previous generation. Subsequently, a strategy is automatically chosen from the corresponding candidate pool to create a mutant vector. In addition, a centroid-based strategy which utilizes the information of multiple superior individuals is designed to balance the population diversity and convergence speed in the second stage. Experiments are conducted on 23 widely used test functions, CEC 2013, and CEC 2014 benchmark sets to demonstrate the performance of the proposed algorithm. The results reveal that the proposed algorithm exhibits better performance compared with several advanced DE variants and some non-DE approaches.

Differential evolution is a fast, robust, and simple population-based stochastic search algorithm for global optimization, which has been widely applied in various fields. However, there are many mutation strategies in DE, which have their own characteristics. Therefore, choosing a best mutation strategy is not easy for a specific problem. Different mutation strategies may be appropriate during different stages of the evolution. In this paper, we propose a DE with multi-stage strategies (DEMS). In DEMS, the evolution process of DE is divided into multiple stages according to the average distance between each individual in the initial population. Each stage has its own strategy candidate pool which includes multiple effective strategies. At the beginning of each generation, the average distance between each individual is first calculated to determine the evolution stage. Then for each target vector in the current population, a mutation strategy is randomly selected from the strategy candidate pool with respect to the stage to produce a offspring vector. Numerical experiments on 15 well-known benchmark functions and the CEC 2015 benchmark sets show that the proposed DEMS is significantly better than, or at least comparable to several state-of-the-art DE variants, in terms of the quality of the final solutions and the convergence rate.

To address the searching problem of protein conformational space in ab-initio protein structure prediction, a novel method using abstract convex underestimation (ACUE) based on the framework of evolutionary algorithm was proposed. Computing such conformations, essential to associate structural and functional information with gene sequences, is challenging due to the high-dimensionality and rugged energy surface of the protein conformational space. As a consequence, the dimension of protein conformational space should be reduced to a proper level. In this paper, the high-dimensionality original conformational space was converted into feature space whose dimension is considerably reduced by feature extraction technique. And, the underestimate space could be constructed according to abstract convex theory. Thus, the entropy effect caused by searching in the high-dimensionality conformational space could be avoided through such conversion. The tight lower bound estimate information was obtained to guide the searching direction, and the invalid searching area in which the global optimal solution is not located could be eliminated in advance. Moreover, instead of expensively calculating the energy of conformations in the original conformational space, the estimate value is employed to judge if the conformation is worth exploring to reduce the evaluation time, thereby making computational cost lower and the searching process more efficient. Additionally, fragment assembly and the Monte Carlo method are combined to generate a series of metastable conformations by sampling in the conformational space. The proposed method provides a novel technique to solve the searching problem of protein conformational space. Twenty small-to-medium structurally diverse proteins were tested, and the proposed ACUE method was compared with It Fix, HEA, Rosetta and the developed method LEDE without underestimate information. Test results show that the ACUE method can more rapidly and more efficiently obtain the near-native protein structure.